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Thursday, December 17, 2015

Via Activation of Oct4
We have a variety of stem cell, progenitor and neuron markers that are often referenced in publication. There represent a qualitative way to determine the state of stem cells as they differentiate.

Here're researchers take Fibroblasts and differentiate them into various progenitors. They were able to further differentiate these progenitors into astrocytes and neurons by: For neuronal differentiation, after 2 days of incubation with 20 nM reversine, cells were then treated with 0.5 μM all-trans-retinoic acid (RA) in serum-free DMEM/F-12 medium supplemented with the ITS for 2 days and switched into serum-free medium in the absence of RA with replacement of the medium every 2-3 days.
Our NSE and GFAP were used to confirm this differentiation.

Wednesday, December 09, 2015

i-Fect TMis a Proven Tool for Gene Manipulation in Studying All Types of Pain

I previously posted on use of our i-Fect Transfection Kit to silence Kv Channels Receptors. This has enabled researchers to study the role of these receptors in vitro and in vivo (see:i-Fect™ Delivers Your siRNA Payload).Sample Data

Image: Mixed neuron-glial cultures stained with Mouse Monoclonal GFAP, and Chicken Polyclonal Neurofilament-NF-L (green). The GFAP antibody stains the network of astrocytes in these cultures, while the NF-L antibody stains neurons and their processes. The blue channel shows the localization of DNA. This antibody also works on formalin fixed paraffin embedded brain tissues. Protocol on Datasheet.

Luc macrophages (CC or BES) were then incorporated into a fl uid mixture containing one of three alternative collagen-based hydrogels, namely, Collagen Hydrogel, Collagen Hydrogel Soft, or Collagen Hydrogel Soft+ (resulting in six possible combinations of macrophages and biopolymer).

Overview of Results

Figure: Bioluminescent imaging of implanted macrophage–biopolymer mixes. IC-21-Luc macrophages (CC or BES) were mixed with biopolymer (Hydrogel, Soft, or Soft+) and subcutaneously injected into the dorsal fl anks of C57Bl/6 mice (three mice per hydrogel, with CC on the left fl ank and BES on the right). Following intraperitoneal injection of delta-luciferin, macrophage bioluminescence was detected using an IVIS Lumina II imaging system. A representative image from day 1 post-implantation (and 25 min post luciferin injection) is shown, indicating the peak detectable radiance (photons s −1 cm −2) in identically sized regions of interest. The CE results were very similar to the BES implants.

Figure: Histological analysis of the macrophage–biopolymer implantation site (MSB staining). At day 4 post-implantation, skin was harvested from each dorsal fl ank, and processed for histology. Where possible, serial sections directly adjacent to those shown in Figure 4 were stained with a modifi ed trichrome stain. Mature fi brillar collagen within the dermis appeared dark blue, while the collagen within the hydrogel was generally lighter blue in appearance, indicating a less highly cross-linked or fi brillar form of collagen. Scale bar: 200 µm.These Collagen Hydrogels are proving easy to use and effective in the hands of our customers. If you are looking for solutions for tissue reconstruction/repair or 3-D in vivo like cell based assays, do not hesitate to contact me @ direct phone: 612-801-1007 or pshuster@neuromics.com. Thank you, Pete Shuster, CEO and Owner, Neuromics.

Thursday, September 17, 2015

Designed for Neuro-muscular Diseases Research
Clients have been using our easy to culture and research proven GFP Labeled Mouse Motor Neurons for Neuro-muscular disease research. This includes the inclusion of the cells in several ALS drug discovery programs being conducted by large Pharma.

Summary: The results demonstrate that, (1) all three commercially available models generated a robust source of proliferating neuroprogenitor cells, and that the assay was sensitive and reproducible when used in a multi-well plate format; (2) there were differences in the response of the rodent and human neuroprogenitor cells to a set of chemicals previously shown to induce apoptosis in vitro; and (3) proliferating neuroprogenitor cells were more sensitive to chemical-induced apoptosis than differentiated neurons, suggesting that neuroprogenitor cells are one of the cell models that should be considered for use in a developmental neurotoxicity screening battery.

Here're more publications of referencing use of hNP1 Neural Progenitors:

Figure: Exogenous CTGF protein increases the number of Sox2-positive cells of a neural progenitor culture. Immunostaining showing Sox2 (A and D) expression of untreated and CTGF-treated cells. C and F show merged pictures of Sox2 immunostaining together with nuclei-DAPI staining. Scale bars 10 μm. G shows the percentage of cells that were positive for Sox2. doi:10.1371/journal.pone.0133689.g002

Friday, August 28, 2015

The roots of Neuromics is providing solutions for the study of nociceptive and neuropathic pain. Our tools are widely used and frequently published.
We are pleased to bring you these small molecules designed to help you better understand the roots of pain and potential therapies.

Figure: Infarct volume after 3 days of reperfusion in the ipsilateral cortex and CP complex in male rats treated with vehicle saline (n=15) or 1 mg/kg per hour BRL 52537 (n=15) started at onset of reperfusion and continued for 22 hours (male-vehicle n=15; male-BRL n=15; mean±SEM). P<.0.05. Related Pub.

Friday, August 14, 2015

Join Neuromics on our‪#‎BrainAdventure‬ to see all the interesting and unique locations the brain ventures. Starting this September, with all orders, you will receive a Neuromics brain keychain. Take a creative picture or short video of the Neuromics Brain Keychain and tweet it @Neuromics, post it on Neuromics Facebook or LinkedIn Page and be entered in our contest. In the meantime, if you want to join the contest, e-mail rose@neuromics.com and she will mail you your brain.

Here researchers study the impact of Marijuana on memory impairment using our one of our 5-HT Serotonin Antibodies: Xavier Viñals, Estefanía Moreno, Laurence Lanfumey, Arnau Cordomí, Antoni Pastor, Rafael de La Torre, Paola Gasperini, Gemma Navarro, Lesley A. Howell, Leonardo Pardo, Carmen Lluís, Enric I. Canela, Peter J. McCormick, Rafael Maldonado, Patricia Robledo. Cognitive Impairment Induced by Delta9-tetrahydrocannabinol Occurs through Heteromers between Cannabinoid CB1 and Serotonin 5-HT2A Receptors. Published: July 9, 2015DOI: 10.1371/journal.pbio.1002194.Impact on Memory: Activation of cannabinoid CB1 receptors (CB1R) by delta9-tetrahydrocannabinol (THC) produces a variety of negative effects with major consequences in cannabis users that constitute important drawbacks for the use of cannabinoids as therapeutic agents. For this reason, there is a tremendous medical interest in harnessing the beneficial effects of THC. Behavioral studies carried out in mice lacking 5-HT2A receptors (5-HT2AR) revealed a remarkable 5-HT2AR-dependent dissociation in the beneficial antinociceptive effects of THC and its detrimental amnesic properties We found that specific effects of THC such as memory deficits, anxiolytic-like effects, and social interaction are under the control of 5-HT2AR, but its acute hypolocomotor, hypothermic, anxiogenic, and antinociceptive effects are not. In biochemical studies, we show that CB1R and 5-HT2AR form heteromers that are expressed and functionally active in specific brain regions involved in memory impairment. Remarkably, our functional data shows that costimulation of both receptors by agonists reduces cell signaling, antagonist binding to one receptor blocks signaling of the interacting receptor, and heteromer formation leads to a switch in G-protein coupling for 5-HT2AR from Gq to Gi proteins. Synthetic peptides with the sequence of transmembrane helices 5 and 6 of CB1R, fused to a cell-penetrating peptide, were able to disrupt receptor heteromerization in vivo, leading to a selective abrogation of memory impairments caused by exposure to THC. These data reveal a novel molecular mechanism for the functional interaction between CB1R and 5-HT2AR mediating cognitive impairment. CB1R-5-HT2AR heteromers are thus good targets to dissociate the cognitive deficits induced by THC from its beneficial antinociceptive properties.

Figures: 5-HT2AR and CB1R form heteromers in transfected cells.
In (A), BRET saturation experiments were performed in HEK-293T cells transfected with 0.025 μg of 5-HT2AR-Rluc cDNA and increasing amounts of CB1R-YFP cDNA (0.05 μg to 1.5 μg, black curve), with 0.5 μg of dopamine D1R-Rluc cDNA and increasing amounts of CB1R-YFP cDNA (0.5 μg to 6 μg, yellow line), or with 0.025 μg of 5-HT2AR-Rluc cDNA and increasing amounts of adenosine A1R-YFP cDNA (0.05 μg to 1.5 μg, red line). The relative amount of BRET is given as a function of 100 x the ratio between the fluorescence of the acceptor (YFP) and the luciferase activity of the donor (Rluc). BRET is expressed as milli BRET units (mBU) and is given as the mean ± standard deviation (SD) of 3–6 experiments grouped as a function of the amount of BRET acceptor. In (B), a schematic representation of fluorescence complementation experiments is depicted in the left panel showing that fluorescence only appears after the YFP Venus hemiprotein complementation due to the proximity of two receptors fused to hemi-YFP Venus proteins (cYFP or nYFP). In the right panel, fluorescence at 530 nm was detected in HEK-293T cells transfected with different amounts of cDNA corresponding to both 5-HT2AR-cYFP and CB1R-nYFP (equal amount for each construct), but not in negative controls in which cells were transfected with cDNA corresponding to 5-HT2AR-cYFP and the noninteracting adenosine A1 receptor-nYFP or CB1R-nYFP and the noninteracting dopamine D1 receptor-cYFP. One-way ANOVA followed by a Dunnett’s multiple comparison test showed a significant fluorescence over basal values in HEK-293T cells (** p< 0.01, *** p< 0.001). In (C), PLAs were performed in HEK-293T cells expressing CB1R and 5-HT2AR. Confocal microscopy images (superimposed sections) are shown in which heteromers appear as green spots. In all cases, cell nuclei were stained with DAPI (blue). Scale bars = 20 μm. In (D), PLAs were performed in nontransfected HEK-293T cells, cells transiently transfected with 0.5 μg of CB1R or 5-HT2AR cDNA (negative controls, white columns), or with increasing amounts of CB1R and 5-HT2AR cDNA (black columns). In each case, the ratio between the number of green spots and the number of cells showing spots (ratio r) was calculated. One-way ANOVA followed by a Dunnett’s multiple comparison test showed a significant PLA staining over nontranfected cells (*** p <; 0.001).
doi:10.1371/journal.pbio.1002194.g004

Yes, memory impairment must be considered as a side effect of Medical Marijuana, I am confident there are human subjects that would volunteer for cognition studies to get a clearer picture of the short and long term impact on users.

Saturday, August 08, 2015

I am pleased by the growing positive response to our Human Endothelial Cells offerings.
Our Human Brain Microvascular Endothelial Cells have, far and away, been the most popular. We want to make it easier for you to "buy and try" We are offering them for only 699 USD/500,000 Cells through Aug. 31st, 2015. Note: customers have passaged these cells up to 15 X.

Monday, August 03, 2015

From Delivery to Stable Expression
Neuromics has a successful track record of helping our clients delivery siRNA, miRNA, Plasmids and other oligos in vitro and in vivo with our Transfection Kits...But my vision with our cell based assay solutions has always been to provide engineered cells and plasmids modified to study your genes of interest. I am pleased to announce we are working with Smart Cell /B-MoGen Technologies to make this happen.
We now can provide:
Gene Transfer and Expression Products
Leveraging the Sleeping Beauty Technology:

Advantages of Sleeping Beauty Transposon System:
· Delivery method is time and cost effective compared to lentiviral delivery.
· Increased cargo-capacity when compared to lentiviral delivery.
· Safest insertion profile of all gene transfer methods.
· Commonly integrated as a single copy.
Custom vector design and assembly, including multi-gene (up to 6) vectors.
We are in the process of formulating standard offerings. In the meantime, I am positioned to offer favorable pricing and terms to early adopters of our Sleeping Beauty Solutions. Please contact me directly pshuster@neuromics.com or 612-801-1007. We can together determine your needs and desired outcomes and provide a statement of work with pricing, project milestones and delivery.

Monday, July 13, 2015

Towards the Development of Disease Specific Assays
Neurodegenerative diseases are becoming increasingly prevalent, especially in the Western societies, with larger percentage of members living to an older age. They have to be seen not only as a health problem, but since they are care-intensive, they also carry a significant economic burden.

Apoptotic pathways are induced in many of these diseases and are key culprits in disease progression.

Figure: Schematic representation of apoptotic pathways. Apoptosis triggered by internal (intrinsic) or external (extrinsic) stress signals that is activated by binding of ligands (e.g. FasL, APO-2L, TRAIL, TNF) to cell surface receptors (e.g. Fas, DR4, DR5, TNF-R1). The intrinsic apoptosis pathway might be triggered by p53 upon DNA damage following exposure to cellular stress. In the intrinsic pathway, death signal reaches mitochondria, leading to release of cytochrome c, which can binds to Apaf1. The cytochrome c/Apaf1 make a complex with pro-caspase-9 (in the presence of dATP), activates caspase-9, which promotes caspase-3 activation, eventually leading to cell death. The extrinsic pathway is initiated through the stimulation of the members of tumor necrosis factor receptor (TNF-R) family (transmembrane death receptors) by their respective ligands. These receptors activate pro-caspases-8, -10 by recruiting the endogenous adaptor protein FADD. Procaspase-8, -10 cleave themselves to form activated caspase-8 or -10. Ultimately, effector enzymes such as caspase-3, -6, -7 are activated in this cascade to mediate apoptosis. Likewise, there can be cross-talk between the intrinsic and extrinsic pathways. For example caspase-8 may cleave Bid to form tBid that is a strong activator of the intrinsic/mitochondrial apoptotic pathway. The intrinsic pathway is usually activated by the recruitment of BAX and BAK to outer mitochondrial membrane, causing cytochrome c release formation of apoptosome and subsequent activation of caspase-9. Activated caspase-9 proteolytically activates caspases-3, -6, and -7. Moreover, some of the effector caspases also can activate caspase-8, forming a positive amplification loop. doi:10.1016/j.pneurobio.2013.10.004.

Tuesday, June 30, 2015

Potent, Pure and Easy to Culture
Assay data continues to roll in. I am pleased by the high lot to lot consistencies of our hN2 Primary Human Neurons resulting in data reproducibility. Here're an examples of outgrowth/tox assays: In conjunction with Molecular Devices' ImageeXpress HCI platform, hN2 differentiated neuronal cells can be used to evaluate potential toxic effects of test compounds on nuerite outgrowth through visualizing cells stained with β-lll tubulin.

Figure: Dose response curves for the effect of neurotoxic agents on neurite outgrowth. hN2™ cells were cultured in the presence of cytotoxic compounds for 72h, stained for β-lll tubulin, imaged with the ImageeXpress and analyzed for neurite outgrowth.
I am always available for product related issues and questions. I cam be reached at direct phone: 612-801-1007 and pshuster@neuromics.com. Pete Shuster, CEO and Owner, Neuromics.

These cells are optimized to provide addition options for in-vitro testing of drug to drug candidate toxicities allowing researchers to rule out the ineffective and potentially toxic small molecules/compounds early in the process.

I want to make it easy for you to buy and try. If you are not delighted with your results, I will replace free of charge or refund your purchase. Please also note the select positive feedback on our other primary and stem cells:We recently used the hMSCs derived from Umbilical Cord Blood. Their performance was nothing less than excellent. We were highly impressed with their morphology and their doubling rate. In addition the cells respond very well to accutase and maintain their performance after passaging. We highly recommend this cell line. Rodney Nash, Ph.D, Georgia State University.Combined Hippocampus, Cortex, and Ventricular Neurons: "I got 10 million cells total after extraction from the tissue. At Day 4 they all developed long axons. Thank you so much for the replacement." Dr. Lidia Gardner, University of Tennessee HSC.Neuromics always provides excellent products and the best customer service. I highly recommend this company's antibodies and neuronal cultures. Kirsten Raehal, Purdue Pharma
I am always available for product related issues and questions. I cam be reached at direct phone: 612-801-1007 and pshuster@neuromics.com. Pete Shuster, CEO and Owner, Neuromics.

Monday, June 22, 2015

Our follow up processes include actively engaging users of our solutions for feedback and input. This includes Rose Ludescher, Manager of Customer Satisfaction, calling or sending e-mails to each user within 3 weeks of shipment. At the core is our product rating website. If users identify products related issues, we replace them or refund their purchases.

We want to hear from you...the good, the bad and the ugly. We welcome opportunities to fix issues.

Image: Mouse striatum stained with D2 cell marker Enkephalin (RA14124) in green and with neuronal marker NeuN in red courtesy of Dr Heike Rebholz of City College of New York.
We are always delighted when we discover our solutions reference in Customer Publications. We try and post all of these to our website. We also post many here in recognition of our customers' research.

For all customers that share data or testimonials, we provide gift cards. It our way of saying thank you, because your feedback matters!